1664results about "Polypeptide with His-tag" patented technology

A recombinant humanized collagen and a preparation method therefor. The recombinant humanized type III collagen can be efficiently and stably expressed in three expression systems of yeast, Escherichia coli, and CHO cells, and has the biological activity superior to that of a commercially available collagen product. The recombinant humanized type III collagen has structural characteristics and functional activity similar to those of a natural collagen, and thus has wide application prospects in biomedical materials, tissue engineering products, cosmetics, foods, health care products or drugs, medical instruments, medical cosmetology and the like.

The invention provides a fusion protein and application thereof in preparation of Brazilian sweet. The fusion protein comprises a tag protein and a Brazilian sweet protein which are connected by a linker; wherein the linker comprises an amino acid sequence as shown in SEQ ID NO: 1, and the amino acid sequence as shown in SEQ ID NO: 1 is directly connected with the Brazilian sweet protein. The invention also provides a preparation method of the Brazilian sweet protein. The preparation method comprises the following steps: treating the fusion protein by using TEV protease and obtaining the Brazilian sweet protein. Large-scale expression of the fusion protein and the brazitame protein is realized by utilizing an escherichia coli expression system, and the obtained brazitame protein can be correctly folded and has high sweetness.

Provided herein are novel immunoglobulin G (IgG) proteases and fusion proteins comprising the same. The IgG proteases find utility in methods of treating diseases mediated in whole or in part by pathogenic immunoglobulin G (IgG) antibodies, including methods for preventing or treating antibody-mediated rejection (AMR) of an organ allograft in an organ-transplant patient.

The invention belongs to the field of biological medicines, and particularly relates to a varicella-zoster virus gE fusion protein as well as a preparation method and application thereof. The varicella-zoster virus gE fusion protein comprises an amino acid sequence as shown in SEQ ID NO: 1 and at least one of the following fragments: a signal peptide, a Foldon sequence, a Ferritin sequence and a His tag sequence, the signal peptide can improve the protein expression quantity, the Foldon sequence can improve the stability of the protein, the particle size of an antigen can be increased, the immunogenicity of the antigen can be improved, and the fusion protein can be used for preparing a medicine for treating the varicella-zoster virus gE fusion protein. The Ferritin sequence can enhance immune response, a vaccine prepared from the gE fusion protein has excellent immunogenicity, and compared with a commercially available vaccine Shingrix, higher humoral immunity and cellular immunity levels can be induced.

The invention relates to the technical field of a preparation method of a type A recombinant botulinum toxin, in particular to a type A botulinum toxin obtained by adopting a gene recombination technology, and the amino acid sequence of the type A botulinum toxin is consistent with the sequence of the type A botulinum toxin prepared by extracting and purifying a wild strain Clostridium botulinum. According to the technical scheme, the single-chain protein of the botulinum toxin A sequentially comprises a botulinum toxin A light chain or a modified botulinum toxin A light chain, a connecting region and a botulinum toxin A heavy chain. According to the scheme, the high-quality recombinant botulinum toxin A is obtained after the single-chain protein is subjected to fermentation, crude purification, enzyme digestion and fine purification. According to the technical scheme, the problems of complex process, long period, high safety risk and the like in the existing process for preparing the botulinum toxin A from the wild bacterial strain clostridium botulinum can be solved, and meanwhile, the problem that the amino acid sequence consistent with that of the botulinum toxin A prepared from the wild bacterial strain clostridium botulinum cannot be obtained through the existing gene recombination technology is solved.

The invention relates to the technical field of recombinant protein expression, in particular to recombinant human-derived III-type collagen as well as a preparation method and application thereof. The invention provides a recombinant human source III type collagen. The amino acid sequence of the recombinant human source III type collagen is shown as SEQ ID NO. 1. The recombinant human-derived III-type collagen prepared by the invention can be better applied to a human body, better exerts biological functions of the collagen, avoids repulsive interaction, is high in safety, can effectively promote cell proliferation and oxidation resistance, and is high in expression yield, stable in batches and good in quality; the recombinant human III type collagen prepared by the invention has the potential of being applied to preparation and production of biological medicines and cosmetics.

The present application discloses a fusion protein comprising a varicella zoster virus protein and a ferritin, a nucleic acid encoding the fusion protein, a pharmaceutical composition comprising the fusion protein, a method of preparing the fusion protein, and a use of the fusion protein.

The invention belongs to the field of biological medicine, and particularly relates to a respiratory syncytial virus F protein mutant and application thereof. The respiratory syncytial virus F protein mutant is characterized in that at least one pair of amino acid residues in an F1 subunit and / or an F2 subunit of F protein polypeptide are substituted by cysteine, and / or an F protein signal peptide part is substituted by a human immunoglobulin light chain signal peptide, and / or alanine A at the 107 site of a multi-base sequence is mutated into arginine R, lysine K at the 134 site is mutated into arginine R, and the amino acid residues in the F1 subunit and / or the F2 subunit of the F protein polypeptide are mutated into arginine. According to the invention, more neutralizing antibody epitopes can be exposed, the stable pre-fusion conformation is maintained, effective neutralizing antibody reaction and antibody binding reaction to the RSV subtypes A and B can be ensured, and the vaccine is suitable for various vaccine forms taking the RSV F protein as the antigen.

The invention discloses a multi-effect repair type collagen and application thereof, and belongs to the technical field of synthetic biology. Transdermal peptide TD-1, a transmembrane region of human XVII type collagen, an extracellular sixteenth non-collagen region, an extracellular fifteenth collagen region and a coding gene of a His tag are selected to be connected in series, and a gene sequence is optimized through pichia pastoris codon selection preference; and then the T-COL17R3 is constructed and expressed to obtain the T-COL17R3. Efficacy experiments prove that compared with similar products sold in the market, T-COL17R3 not only has excellent transdermal performance, but also has better cell proliferation and migration promoting capability, better capability of resisting early saccharification product ketoamine and later saccharification product dicarbonyl compounds, better capability of scavenging free radicals and resisting elastase, and can be used for preparing the anti-epidermal drug. In addition, synthesis of melanin in B16-F10 cells can be reduced, so that T-COL17R3 has excellent application potential in development of pharmaceutical compositions or skin care products with repairing, anti-wrinkle and firming or whitening and brightening effects.

A recombinant humanized collagen type I alpha-1 (rhCol1A1), and an expression vector and use thereof are provided, belonging to the technical field of bioengineering. The rhCol1A1 has an amino acid sequence of SEQ ID NO: 1. This aim is to solve the problem of difficulty in expressing soluble full-length humanized collagen in Escherichia coli prokaryotic expression systems, and differences between the recombinant collagen and humanized collagen in yeast expression systems. It is proposed that the rhCol1A1 chain is expressed in Expi293F human embryonic kidney cells, and the protein expression level is increased using a fusion expression plasmid, enabling the expression of full-length humanized collagen type I alpha 1 chain. Activity studies have shown that the expressed rhCol1A1 exhibits a desirable activity in promoting cell migration, and may be widely used in the fields of biomedicine, tissue engineering, skin care, and food.

Provided herein are albumin binding proteins and methods of making and using thereof.

The invention specifically discloses a composition for detecting NAD < + > and application thereof, and relates to the technical field of biological detection. According to the composition for detecting NAD < + >, the composition comprises an NAD < + > binding protein fragment A connected with an Fc tag, an NAD < + > binding protein fragment B and a connecting peptide, the detection result is accurate and reliable, and meanwhile the technical difficulty, uncertainty and high cost faced by antibody screening in a traditional method are avoided.

The invention relates to the technical field of biology, in particular to a pseudorabies virus gD recombinant protein, a subunit vaccine and a preparation method and application thereof.The gD recombinant protein sequentially comprises a signal peptide, a connecting peptide, the 21st-350th site of the gD protein, an enzyme cutting site and a His tag from the N end to the C end, the amino acid sequence of the 21st-350th site of the gD protein is shown as SEQ ID No.4, and the amino acid sequence of the 21st-350th site of the gD protein is shown as SEQ ID No.6. The pseudorabies virus gD recombinant protein has high immunogenicity, the prepared subunit vaccine can generate a specific antibody with neutralizing activity, mice do not die after challenge, the protection rate is 100%, the protection rate after challenge of pigs is 100%, pathological sections only show slight pathological changes, and the pseudorabies virus gD recombinant protein has wide application prospects.

Provided herein are albumin binding proteins and methods of making and using thereof.

The present invention provides an antibody binding to human CD73, a preparation method therefor, and a use thereof. The monoclonal antibody of the present invention can bind to a CD73 antigen with high specificity, has high affinity, has remarkable anti-tumor activity and the like, and has excellent clinical application prospects.

The present invention relates to anti-LYPD3 antibodies that bind to LYPD3 in an O-glycosylation dependent manner. Therefore, the antibody has specificity to tumor-associated LYPD3 (LYPD3). The invention also provides a pharmaceutical composition containing the anti-LYPD3 antibody and an application of the pharmaceutical composition in cancer treatment.

The present invention relates to a fusion protein that produces point mutations in cells, its preparation and use. Specifically, the fusion protein provided by the present invention contains a Cas enzyme that lacks cytosine deaminase and nuclease activity and retains helicase activity, or is formed by a Cas enzyme that lacks cytosine deaminase and nuclease activity and retains helicase activity. The present invention also relates to the coding sequence of the fusion protein, a polynucleotide sequence containing the coding sequence, a nucleic acid construct containing the polynucleotide sequence, a corresponding host cell, a method for producing point mutations in a cell, and a kit, etc. By using the present invention, it is possible to achieve site-directed mutagenesis while obtaining high mutation efficiency and a variety of mutation combinations in a specific gene region.

Fungal sweet-taste modifying protein variants (designated Honey Truffle Sweetener (HTS) protein variants), and polynucleotides encoding the protein variants are described. Specifically, Myd (HTS) protein variants with sweet taste modulation activity, and the cDNA encoding the same, are described, along with methods for isolating such cDNA and for isolating and expressing such proteins. Also disclosed are sweetening compositions which include the proteins of the invention, and methods to provide improved flavor to a product for oral administration.

The invention discloses a haemonchus contortus subunit vaccine, and belongs to the field of veterinary vaccines. The subunit vaccine comprises an antigen protein MEP3, an antigen protein AP1, an antigen protein AP8, an antigen protein H11-2, an antigen protein H11-4 and an antigen protein AP5 of galactosylated and modified haemonchus contortus, and the antigen protein MEP3, the antigen protein AP1, the antigen protein AP8, the antigen protein H11-2, the antigen protein H11-4 and the antigen protein AP5 of galactosylated and modified haemonchus contortus. The galactosylation modification of the antigen protein is realized by jointly expressing glycosyl transferase HcGALT of haemonchus contortus and one or more of the proteins in eukaryotic cells. The antigen protein modified by galactosylation in the subunit vaccine is closer to the glycosylation modification of natural protein, and can provide effective immune protection after the goat is infected with haemonchus contortus, and the egg laying rate is reduced by 81.43%. The invention lays a foundation for development of haemonchus contortus subunit vaccines, provides an effective technical means for prevention and treatment of the haemonchus contortus subunit vaccines, and has important application value and popularization prospect.

Provided is a low molecular weight collagen with a 164.88° triple helix structure. Relates to a small molecule collagen, a preparation method and use thereof. The collagen of the present application comprises an amino acid sequence as shown in SEQ ID NO:1 or a variant amino acid sequence after mutation of the amino acid sequence, and the variant amino acid sequence retains the function of the amino acid sequence as shown in SEQ ID NO:1. The amino acid sequence of the collagen of the present application is shorter and has better transdermal absorption performance.

Provided is a method for biosynthesis of a human body structural material type-VIII collagen. Also provided is a collagen, comprising one or more repeating units, wherein the repeating units are connected directly or by means of linkers, and each repeating unit comprises an amino acid sequence selected from the group consisting of SEQ ID NO: 1, 4, 7, 10, 13, 16, 19, 22, 25 or 28, or a variant thereof. The collagen can be used for biological dressings, human body bionic materials or plastic surgery materials. According to the method, the type-VIII collagen is produced by utilizing a genetic engineering technology, thereby overcoming the defects in the prior art.

The present invention relates to modified Dda helicases which can be used to control the movement of analytes such as polynucleotides. The modified Dda helicases are used in analyte detection and characterisation. The present invention also relates to novel protein pores and their uses in analyte detection and characterisation. The invention particularly relates to an isolated pore complex formed by a CsgG-like pore and a modified CsgF peptide, or a homologue or mutant thereof, thereby incorporating an additional channel constriction or reader head in the nanopore.

The invention discloses a T7 RNA polymerase mutant capable of being used for preparing mRNA vaccines and improving thermal stability and transcription performance and a preparation method of the T7 RNA polymerase mutant, relates to the technical field of biology, and particularly discloses a T7 RNA polymerase mutant containing mutation sites K8G, H27K, A70N, K180D and S192P / S192G relative to wild type T7 RNA polymerase. The amino acid sequence of the wild type T7 RNA polymerase is as shown in SEQ ID NO: 1. Compared with wild type T7 RNA polymerase, the T7 RNA polymerase has higher specific enzyme activity and heat stability, the heat resistance of one mutant is improved, a large number of transcription products can still be obtained at 45 DEG C, and transcription by-products of the other mutant are reduced. The mutant T7 RNA polymerase disclosed by the invention has a good application prospect in the fields of RNA vaccines and medicines, in-vitro transcription, gene editing and the like.

Disclosed herein are monospecific and multi-specific antibodies with specificities for one or more of OX40, CD40, 4-1BB, HSA, IL-22, or EGFR. Specifically, the disclosure provides variable heavy (VHH) domains of the antibodies with specificities for one or more of OX40, CD40, 4-1BB, HSA, IL-22, or EGFR, and associated sequences. Further discloses are methods of using the monospecific or multi-specific antibodies for treating ah autoimmune disease.

Relating to the field of biotechnology, disclosed are a recombinant human elastin peptide with anti-aging effects and a use thereof. The recombinant human elastin peptide comprises an amino acid sequence shown in SEQ ID NO:1 or a fragment of the amino acid sequence shown in SEQ ID NO:1, the fragment comprises 30% or more of SEQ ID NO:1, or a mutant of the amino acid sequence shown in SEQ ID NO:1 or the fragment of the amino acid sequence shown in SEQ ID NO:1. A disclosed method for producing elastin peptides by using a yeast expression system enables the targeted synthesis of highly effective protein fragments with 100% homology to human proteins, while achieving low costs and facilitating large-scale production, avoiding animal-derived sources and viral risks. This makes the recombinant human elastin peptide safe for use as an anti-aging and anti-photoaging active ingredient in cosmetics, while also supporting a use thereof in the field of biomedical materials.

The present invention provides compositions and methods comprising an activator of interleukin-18 (IL-18) activity for use in therapeutic and non-therapeutic applications. The activator provides IL-18 signaling activity even in the presence of an inhibitory molecule 5 such as IL-18 binding protein (IL-18BP).

To provide antioxidant peptides derived from Xuanwei ham, and to provide methods for preparation and activity measurement thereof.SOLUTION: The present invention provides an antioxidant peptide with a specific amino acid sequence derived from Xuanwei ham. The present invention uses a rigid linking peptide and a flexible linking peptide to link a hexapeptide derived from Xuanwei ham, growing the peptide chain to make it suitable for genetic recombinant expression, while increasing the proportion of a target polypeptide in a recombinant protein and significantly increasing the expression level. The rigid linking peptide has an α-helical structure, which makes it easier for the antioxidant peptide to form a stable secondary structure after serial linking, and can effectively separate the active functional domain.The flexible linking peptide is mainly composed of glycine Gly, and since it has a low molecular weight, the flexibility of a polypeptide backbone structure is ensured.SELECTED DRAWING: Figure 1

The present invention provides antibodies that bind to and stabilize human Triggering Receptor Expressed on Myeloid cells 2 (TREM2) protein and methods of using these antibodies.

The present application discloses a T7 RNA polymerase mutant with improved thermostability and transcription performance that can be used to prepare an mRNA vaccine and a preparation method thereof, and relates to the field of biotechnology. The present invention specifically discloses a T7 RNA polymerase mutant containing mutation sites K8G, H27K, A70N, K180D, and S192P / S192G relative to the wild-type T7 RNA polymerase; the amino acid sequence of the wild-type T7 RNA polymerase is shown in SEQ ID NO: 1. Compared with the wild-type T7 RNA polymerase, it has higher specific enzyme activity and thermal stability, wherein one mutant has improved heat resistance, and a large amount of transcription products can still be obtained at 45°C, and the transcription byproducts of another mutant are reduced. The mutant T7 RNA polymerase disclosed in the present invention has good application prospects in the fields of RNA vaccines and drugs, in vitro transcription, gene editing, etc.

Provided are a recombinant human interleukin-11 mutant, and a conjugate of the mutant conjugated to a chemical molecule and a use thereof. The recombinant human interleukin-11 mutant achieves soluble expression in an Escherichia coli expression system by means of an SUMO sequence tandem fusion method. The conjugate is obtained by the site-specific conjugation of the mutant to the chemical molecule at a mutant amino acid residue in position W147 by means of a covalent bond. The chemical molecule is at least one of polyethylene glycol, an alkanoic acid aliphatic chain, and a hydrophilic high-molecular polymer containing an alkanoic acid aliphatic chain. The conjugate retains the binding capacity to human interleukin-11 receptor A while demonstrating significantly reduced binding capacity to human IL-6 signal transducer receptor. The conjugate shows the ability to block the signaling between human IL-11 and IL-11 RA and between human IL-11 and GP130 either in vivo or in vitro and can be used for the treatment of associated diseases with tissue fibrosis as a clinical manifestation caused by elevated primary or secondary IL-11.